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Team:UNIPV-Pavia/Notebook/Week16
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!|[[Team:UNIPV-Pavia/Notebook/Week20|Week 20]] | !|[[Team:UNIPV-Pavia/Notebook/Week20|Week 20]] | ||
!|[[Team:UNIPV-Pavia/Notebook/Week21|Week 21]] | !|[[Team:UNIPV-Pavia/Notebook/Week21|Week 21]] | ||
| + | |- | ||
| + | !|[[Team:UNIPV-Pavia/Notebook/Week22|Week 22]] | ||
| + | !|[[Team:UNIPV-Pavia/Notebook/Week23|Week 23]] | ||
| + | !|[[Team:UNIPV-Pavia/Notebook/Week24|Week 24]] | ||
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<br> | <br> | ||
*We had to perform these ligations: | *We had to perform these ligations: | ||
| - | **22- | + | **Lig.22-R0062 |
| - | **31-22 | + | **Lig.31-Lig.22 |
| - | *but we didn't have enough | + | *but we didn't have enough Lig.22 and Lig.31 plasmids... |
| - | *We infected 9 ml of LB + Amp with 30 µl of 22 and 31 glycerol stocks. | + | *We infected 9 ml of LB + Amp with 30 µl of Lig.22 and Lig.31 glycerol stocks. |
'''09/2/08''' | '''09/2/08''' | ||
<br> | <br> | ||
| - | *Glycerol stocks/miniprep for 22 and 31. | + | *Glycerol stocks/miniprep for Lig.22 and Lig.31. |
*Plasmid digestion for: | *Plasmid digestion for: | ||
| - | **22 (E-S) | + | **Lig.22 (E-S) |
| - | **22 (X-P) | + | **Lig.22 (X-P) |
| - | **31 (S-P) | + | **Lig.31 (S-P) |
| - | ** | + | **R0062 (E-X) |
*Run/gel extraction. | *Run/gel extraction. | ||
*Ligations: | *Ligations: | ||
| - | **31-22 (= | + | **Lig.31-Lig.22 (=Lig.35) |
| - | **22- | + | **Lig.22-R0062 (=Lig.30) |
| - | *We incubated ligations at 16°C | + | *We incubated ligations at 16°C overnight. |
'''09/3/08''' | '''09/3/08''' | ||
| Line 84: | Line 88: | ||
'''09/4/08''' | '''09/4/08''' | ||
<br> | <br> | ||
| - | *Colony PCR for the two ligation plates: | + | *Colony PCR for the two ligation plates |
| + | |||
| + | {| | ||
| + | |[[Image:pv_colonypcr_30_35again.jpg|thumb|370px|left|Marker 1Kb, Lig.30 (5 colonies), Marker 1Kb, Lig.35 (5 colonies)]] | ||
| + | |} | ||
*Gel results: | *Gel results: | ||
| - | **30 (3rd colony) | + | **Lig.30 (3rd colony) |
| - | **35 (gel showed an unexpected contaminant...we decided to re-perform this ligation) | + | **Lig.35 (gel showed an unexpected contaminant...we decided to re-perform this ligation) |
'''09/5/08''' | '''09/5/08''' | ||
<br> | <br> | ||
| - | *Glycerol stock/miniprep for 30. | + | *Glycerol stock/miniprep for Lig.30. |
| - | *We sent 30( | + | *We sent Lig.30 (3rd col) to Primm for sequencing. |
Latest revision as of 21:27, 26 October 2008
 Home
|  The Team
|  The Project
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|  Parts Submitted to the Registry
|
|---|---|---|---|---|
 Dry Lab
|  Wet Lab
|  Modeling
|  Protocols
|  Activity Notebook
|
Notebook
| Week 1 | Week 2 | Week 3 | Week 4 | Week 5 | Week 6 | Week 7 |
|---|---|---|---|---|---|---|
| Week 8 | Week 9 | Week 10 | Week 11 | Week 12 | Week 13 | Week 14 |
| Week 15 | Week 16 | Week 17 | Week 18 | Week 19 | Week 20 | Week 21 |
| Week 22 | Week 23 | Week 24 |
Week 16: 09/1/08 - 09/5/08
09/1/08
- We had to perform these ligations:
- Lig.22-R0062
- Lig.31-Lig.22
- but we didn't have enough Lig.22 and Lig.31 plasmids...
- We infected 9 ml of LB + Amp with 30 µl of Lig.22 and Lig.31 glycerol stocks.
09/2/08
- Glycerol stocks/miniprep for Lig.22 and Lig.31.
- Plasmid digestion for:
- Lig.22 (E-S)
- Lig.22 (X-P)
- Lig.31 (S-P)
- R0062 (E-X)
- Run/gel extraction.
- Ligations:
- Lig.31-Lig.22 (=Lig.35)
- Lig.22-R0062 (=Lig.30)
- We incubated ligations at 16°C overnight.
09/3/08
- We transformed/plated ligations.
09/4/08
- Colony PCR for the two ligation plates
 |
- Gel results:
- Lig.30 (3rd colony)
- Lig.35 (gel showed an unexpected contaminant...we decided to re-perform this ligation)
09/5/08
- Glycerol stock/miniprep for Lig.30.
- We sent Lig.30 (3rd col) to Primm for sequencing.
